Inbred corn line G06-NP2546

ABSTRACT

Broadly, this invention provides for an inbred corn line designated G06-NP2546, methods for producing a corn plant by crossing plants of the inbred line G06-NP2546 with plants of another corn plants. The invention relates to the various parts of inbred G06-NP2546 including culturable cells. This invention also relates to methods for introducing transgenic transgenes into inbred corn line G06-NP2546 and plants produced by said methods.

FIELD OF THE INVENTION

This invention is in the field of corn breeding, specifically relatingto an inbred corn line designated G06-NP2546. This invention also is inthe field of hybrid maize production employing the present inbred.

BACKGROUND OF THE INVENTION

The original maize plant was indigenous to the Western Hemisphere. Theplants were weedlike and only through the efforts of early breeders werecultivated crop species developed. The crop cultivated by earlybreeders, like the crop today, could be wind pollinated. The physicaltraits of maize are such that wind pollination results inself-pollination or cross-pollination between plants. Each maize planthas a separate male and female flower that contributes to pollination,the tassel and ear, respectively. Natural pollination occurs when windtransfers pollen from tassel to the silks on the corn ears. This type ofpollination has contributed to the wide variation of maize varietiespresent in the Western Hemisphere.

The development of a planned breeding program for maize only occurred inthe last century. A large part of the development of the maize productinto a profitable agricultural crop was due to the work done by landgrant colleges. Originally, maize was an open pollinated variety havingheterogeneous genotypes. The maize farmer selected uniform ears from theyield of these genotypes and preserved them for planting the nextseason. The result was a field of maize plants that were segregating fora variety of traits. This type of maize selection led to, at most,incremental increases in seed yield.

Large increases in seed yield were due to the work done by land grantcolleges that resulted in the development of numerous hybrid cornvarieties in planned breeding programs. Hybrids were developed byselecting corn lines and selfing these lines for several generations todevelop homozygous pure inbred lines. One selected inbred line wascrossed with another selected inbred line to produce hybrid progeny(F1). The resulting hybrids, due to heterosis, are robust and vigorousplants. Inbreds on the other hand are mostly homozygous. Thishomozygosity renders the inbred lines less vigorous. Inbred seed can bedifficult to produce since the inbreeding process in corn linesdecreases the vigor. However, when two inbred lines are crossed, thehybrid plant evidences greatly increased vigor and seed yield comparedto open pollinated, segregating maize plants. An important consequenceof the homozygosity and the homogenity of the inbred maize lines is thatall hybrid seed produced from any cross of two such elite lines will bethe same hybrid seed and make the same hybrid plant. Thus the use ofinbreds makes hybrid seed which can be reproduced readily. The hybridplant in contrast does not produce hybrid seed that is readilyreproducible. The seed on a hybrid plant is segregating for traits.

The ultimate objective of the commercial maize seed companies is toproduce high yielding, agronomically sound plants that perform well incertain regions or areas of the Corn Belt. To produce these types ofhybrids, the companies must develop inbreds, which carry needed traitsinto the hybrid combination. Hybrids are not often uniformly adapted forthe entire Corn Belt, but most often are specifically adapted forregions of the Corn Belt. Northern regions of the Corn Belt requireshorter season hybrids than do southern regions of the Corn Belt.Hybrids that grow well in Colorado and Nebraska soils may not flourishin richer Illinois and Iowa soil. Thus, a variety of major agronomictraits is important in hybrid combination for the various Corn Beltregions, and has an impact on hybrid performance.

Inbred line development and hybrid testing have been emphasized in thepast half-century in commercial maize production as a means to increasehybrid performance. Inbred development is usually done by pedigreeselection. Pedigree selection can be selection in an F₂ populationproduced from a planned cross of two genotypes (often elite inbredlines), or selection of progeny of synthetic varieties, open pollinated,composite, or backcrossed populations. This type of selection iseffective for highly inheritable traits, but other traits, for example,yield requires replicated test crosses at a variety of stages foraccurate selection.

Maize breeders select for a variety of traits in inbreds that impacthybrid performance along with selecting for acceptable parental traits.Such traits include: yield potential in hybrid combination; dry down;maturity; grain moisture at harvest; greensnap; resistance to rootlodging; resistance to stalk lodging; grain quality; disease and insectresistance; ear and plant height. Additionally, Hybrid performance willdiffer in different soil types such as low levels of organic matter,clay, sand, black, high pH, low pH; or in different environments such aswet environments, drought environments, and no tillage conditions. Thesetraits appear to be governed by a complex genetic system that makesselection and breeding of an inbred line extremely difficult. Even if aninbred in hybrid combination has excellent yield (a desiredcharacteristic), it may not be useful because it fails to haveacceptable parental traits such as seed yield, seed size, pollenproduction, good silks, plant height, etc.

To illustrate the difficulty of breeding and developing inbred lines,the following example is given. Two inbreds compared for similarity of29 traits differed significantly for 18 traits between the two lines. If18 simply inherited single gene traits were polymorphic with genefrequencies of 0.5 in the parental lines, and assuming independentsegregation (as would essentially be the case if each trait resided on adifferent chromosome arm), then the specific combination of these traitsas embodied in an inbred would only be expected to become fixed at arate of one in 262,144 possible homozygous genetic combinations.Selection of the specific inbred combination is also influenced by thespecific selection environment on many of these 18 traits which makesthe probability of obtaining this one inbred even more remote. Inaddition, most traits in the corn genome are regrettably not singledominant genes but are multi-genetic with additive gene action notdominant gene action. Thus, the general procedure of producing a nonsegregating F₁ generation and self pollinating to produce a F₂generation that segregates for traits and selecting progeny with thevisual traits desired does not easily lead to a useful inbred. Greatcare and breeder expertise must be used in selection of breedingmaterial to continue to increase yield and the agronomics of inbreds andresultant commercial hybrids.

Certain regions of the Corn Belt have specific difficulties that otherregions may not have. Thus the hybrids developed from the inbreds haveto have traits that overcome or at least minimize these regional growingproblems. Examples of these problems include in the eastern corn beltGray Leaf Spot, in the north cool temperatures during seedlingemergence, in the Nebraska region CLN (corn Lethal necrosis and in thewest soil that has excessively high pH levels. The industry oftentargets inbreds that address these issues specifically forming nicheproducts. However, the aim of most large seed producers is to provide anumber of traits to each inbred so that the corresponding hybrid canuseful in a broader regions of the Corn Belt. The new biotechnologytechniques such as Microsatellites, RFLPs, RAPDs and the like haveprovided breeders with additional tools to accomplish these goals.

SUMMARY OF THE INVENTION

The present invention relates to an inbred corn line G06-NP2546.Specifically, this invention relates to plants and seeds of this line.Additionally, this relates to a method of producing from this inbred,hybrid seed corn and hybrid plants with seeds from such hybrid seed.More particularly, this invention relates to the unique combination oftraits that combine in corn line G06-NP2546.

Generally then, broadly the present invention includes an inbred cornseed designated G06-NP2546. This seed produces a corn plant.

The invention also includes the tissue culture of regenerable cells ofG06-NP2546 wherein the cells of the tissue culture regenerates plantscapable of expressing the genotype of G06-NP2546. The tissue culture isselected from the group consisting of leaf, pollen, embryo, root, roottip, guard cell, ovule, seed, anther, silk, flower, kernel, ear, cob,husk and stalk, cell and protoplast thereof. The corn plant regeneratedfrom G06-NP2546 or any part thereof is included in the presentinvention. The present invention includes regenerated corn plants thatare capable of expressing G06-NP2546's genotype, phenotype or mutants orvariants thereof.

The invention extends to hybrid seed produced by planting, inpollinating proximity which includes using preserved maize pollen asexplained in U.S. Pat. No. 5,596,838 to Greaves, seeds of corn inbredlines G06-NP2546 and another inbred line if preserved pollen is notused; cultivating corn plants resulting from said planting; preventingpollen production by the plants of one of the inbred lines if two areemployed; allowing cross pollination to occur between said inbred lines;and harvesting seeds produced on plants of the selected inbred. Thehybrid seed produced by hybrid combination of plants of inbred corn seeddesignated G06-NP2546 and plants of another inbred line are apart of thepresent invention. This inventions scope covers hybrid plants and theplant parts including the grain and pollen grown from this hybrid seed.

The invention further includes a method of hybrid F1 production. A firstgeneration (F1) hybrid corn plant produced by the process of plantingseeds of corn inbred line G06-NP2546; cultivating corn plants resultingfrom said planting; permitting pollen from another inbred line to crosspollinate inbred line G06-NP2546; harvesting seeds produced on plants ofthe inbred; and growing a harvested seed are part of the method of thisinvention.

Likewise included is a first generation (F1) hybrid corn plant producedby the process of planting seeds of corn inbred line G06-NP2546;cultivating corn plants resulting from said planting; permitting pollenfrom inbred line G06-NP2546 to cross pollinate another inbred line;harvesting seeds produced on plants of the inbred; and growing a plantfrom such a harvested seed.

The inbred corn line G06-NP2546 and at least one transgenic gene adaptedto give G06-NP2546 additional and/or altered phenotypic traits arewithin the scope of the invention. Such transgenes are usuallyassociated with regulatory elements (promoters, enhancers, terminatorsand the like). Presently, trangenes provide the invention with traitssuch as insect resistance, herbicide resistance, disease resistanceincreased or deceased starch or sugars or oils, increased or decreasedlife cycle or other altered trait.

The present invention includes inbred corn line G06-NP2546 and at leastone transgenic gene adapted to give G06-NP2546 modified starch traits.Furthermore this invention includes the inbred corn line G06-NP2546 andat least one mutant gene adapted to give modified starch, acid or oiltraits. The present invention includes the inbred corn line G06-NP2546and at least one transgenic gene for example: VIP, bacillusthuringiensis, the bar or pat gene encoding Phosphinothricin acetylTransferase, Gdha gene, GOX, EPSP synthase gene, low phytic acidproducing gene, and zein. The inbred corn line G06-NP2546 and at leastone transgenic gene useful as a selectable marker or a screenable markeris covered by the present invention.

A tissue culture of the regenerable cells of hybrid plants produced withuse of G06-NP2546 genetic material is covered by this invention. Atissue culture of the regenerable cells of the corn plant produced bythe method described above is also included.

Definitions

In the description and examples, which follow, a number of terms areused. In order to provide a clear and consistent understanding of thespecifications and claims, including the scope to be given such terms,the following definitions are provided.

Color Choices:  1. light green  2. medium green  3. dark green  4. verydark green  5. green-yellow  6. pale yellow  7. yellow  8. yelow-orange 9. salmon 10. pink-orange 11. pink 12. light red 13. cherry red 14. red15. red and white 16. pale purple (describe) 17. purple 18. colorless19. white 20. white capped 21. buff 22. tan 23. brown 24. bronze 25.variegated 26. other (describe)

Form Input # ABR. Description Value A1 EMRGN Final number of plants perplot # A2 REGNN Region Developed: 1. Northwest # 2. Northcentral 3.Northeast 4. Southeast 5. Southcentral 6. Southwest 7. Other A3 CRTYNCross type: 1. sc 2. dc 3. 3w 4. msc # 5. m3w 6. inbred 7. rel. line 8.other A4 KRTPN Kernel type: 1. sweet 2. dent 3. flint # 4. flour 5. pop6. ornamental 7. pipecorn 8. other A5 EMERN Days to Emergence EMERN #Days B1 ERTLP % Root lodging: (before anthesis): # % B2 GRSNP % Brittlesnapping: (before anthesis): # % C1 TBANN Tassel branch angle of 2ndprimary lateral degree branch (at anthesis): C10 HUPSN Heat units to 50%pollen shed: (from # HU emergence) C11 SLKCN Silk color: #/Munsell valueC12 HU5SN Heat units to 50% silk: (from emergence) # HU C13 DSAZN Daysto 50% silk in adapted zone: # Days C14 HU9PN Heat units to 90% pollenshed: (from # HU emergence) C15 HU19N Heat units from 10% to 90% pollenshed: # HU C16 DA19N Days from 10% to 90% pollen shed: # Days C2 LSPURLeaf sheath pubescence of second leaf # above the ear (at anthesis) 1-9(1 = none): C3 ANGBN Angle between stalk and 2nd leaf above degree theear (at anthesis): C4 CR2LN Color of 2nd leaf above the ear (at#/Munsell anthesis): value C5 GLCRN Glume Color: #/Munsell value C6GLCBN Glume color bars perpendicular to their # veins (glume bands): 1.absent 2. present C7 ANTCN Anther color: #/Munsell value C8 PLQUR PollenShed: 1-9 (0 = male sterile) # C9 HU1PN Heat units to 10% pollen shed: #HU (from emergence) D1 LAERN Number of leaves above the top ear node: #D10 LTBRN Number of lateral tassel branches that # originate from thecentral spike: D11 EARPN Number of ears per stalk: # D12 APBRRAnthocyanin pigment of brace roots: # 1. absent 2. faint 3. moderate 4.dark D13 TILLN Number of tillers: # D14 HSKCN Husk color 25 days after50% silk: (fresh) #/Munsell value D2 MLWVR Leaf marginal waves: 1-9 (1 =none) # D3 LFLCR Leaf longitudinal creases: 1-9 (1 = none) # D4 ERLLNLength of ear leaf at the top ear node: # cm D5 ERLWN Width of ear leafat the top ear node at the # cm widest point: D6 PLHTN Plant height totassel tip: # cm D7 ERHCN Plant height to the top ear node: # cm D8LTEIN Length of the internode between the ear # cm node and the nodeabove: D9 LTASN Length of the tassel from top leaf collar to # cm tasseltip: E1 HSKDN Husk color 65 days after 50% silk: (dry) #/Munsell valueE10 DSGMN Days from 50% silk to 25% grain moisture # Days in adaptedzone: E11 SHLNN Shank length: # cm E12 ERLNN Ear length: # cm E13 ERDINDiameter of the ear at the midpoint: # mm E14 EWGTN Weight of a huskedear: # gm E15 KRRWR Kernel rows: 1. indistinct 2. distinct E16 KRNARKernel row alignment: 1. straight # 2. slightly curved 3. curved E17ETAPR Ear taper: 1. slight 2. average 3. extreme # E18 KRRWN Number ofkernel rows: # E19 COBCN Cob color: #/Munsell value E2 HSKTR Husktightness 65 days after 50% silk: 1-9 # (1 = loose) E20 COBDN Diameterof the cob at the midpoint: # mm E21 YBUAN Yield: # kg/ha E22 KRTENEndosperm type: 1. sweet 2. extra sweet 3 3. normal 4. high amylose 5.waxy 6. high protein 7. high lysine 8. super sweet 9. high oil 10. otherE23 KRCLN Hard endosperm color: #/Munsell value E24 ALECN Aleuronecolor: #/Munsell value E25 ALCPR Aleurone color pattern: 1. homozygous #2. segregating E26 KRLNN Kernel length: # mm E27 KRWDN Kernel width: #mm E28 KRDPN Kernel thickness: # mm E29 K1KHN 100 kernel weight: # gm E3HSKCR Husk extension: 1. short (ear exposed) # 2. medium (8 cm) 3. long(8-10 cm) 4. very long (>10 cm) E30 KRPRN % round kernels on 13/64slotted screen: # % E4 HEPSR Position of ear 65 days after 50% silk:# 1. upright 2. horizontal 3. pendent E5 STGRP Staygreen 65 days afteranthesis: 1-9 # (1 = worst) E6 DPOPP % dropped ears 65 days afteranthesis: % E7 LRTRP % root lodging 65 days after anthesis: % E8 HU25NHeat units to 25% grain moisture: (from # HU emergence) E9 HUSGN Heatunits from 50% silk to 25% grain # HU moisture in adapted zone:

DETAILED DESCRIPTION OF THE INVENTION

G06-NP2546 is shown in comparison with Mo17 a standard used forcomparison by the US PVP office.

The inbred provides uniformity and stability within the limits ofenvironmental influence for traits as described in the VarietyDescription Information (Table 1) that follows.

The inbred has been self-pollinated for a sufficient number ofgenerations to give inbred uniformity. During plant selection in eachgeneration, the uniformity of plant type was selected to ensurehomozygosity and phenotypic stability. The line has been increased inisolated farmland environments with data on uniformity and agronomictraits being observed to assure uniformity and stability. No varianttraits have been observed or are expected in G06-NP2546.

The best method of producing the invention, G06-NP2546 which issubstantially homozygous, is by planting the seed of G06-NP2546 which issubstantially homozygous and self-pollinating or sib pollinating theresultant plant in an isolated environment, and harvesting the resultantseed.

TABLE 1 G06-NP2546 VARIETY DESCRIPTION INFORMATION #1 Type: corn - Dent#2 Region where developed in the USA Southcentral #3 Maturity Days HeatUnits 59 1222.5 from emergence to 50% of plants in silk 59 1221.4 fromemergence to 50% plants in pollen    81 from 10% to 90% of pollen shed#4 Plant Traits Plant height 210.8 cm. Ear height  74.5 cm Length of topear internode  16.1 cm Average no. Tillers 0.1 Average no. of ears perstalk 1.0 Anthocyanin of brace roots faint #5 Leaf Traits Width of earnode leaf  9.2 cm Length of ear node leaf  79.0 cm No. of leaves abovetop ear 005 Degrees of leaf angle  18 Leaf color 04 (Very Dark green)(Munsell Code 5GY 3/4) Leaf sheath pubescence  6* Marginal waves  5**Longitudinal creases  6** #6 Tassel Traits No. of primary lateralbranches 03 Branch angle from central spike 20 Tassel length  40.4 cmPollen shed  5*** Anther Color 14 (red) (Munsell Code 5R 4/4) GlumeColor 26 medium green (Munsell Code 5GY 6/6) Bar glumes present #7a Ear(unhusked) Traits Silk Color  5 Green-yellow (Munsell Code 2.5GY 8/8)Fresh husk color  5 Green-yellow (Munsell Code 5GY 7/6) Dry husk color22 tan (Munsell Code 2.5Y 8/4) Position of ear at dry husk stagehorizontal Husk tightness 6**** Husk extension  2 short (meduim) #7b Ear(husked) Traits Ear length  14.3 cm Ear diameter at midpoint  44.4 mmEar weight 127.5 gm No. of kernel rows 13 Kernel rows Distinct Rowalignment slightly curved Shank length  6.4 cm Ear taper average #8Kernel (dried) Kernel length  11.2 mm Kernel width  9.2 mm Kernelthickness  4.0 mm % round kernels 34.1 Aleruone color pattern HomozygousAleurone color white Hard endosperm color 7 yellow (Munsell Code 2.5Y8/10) Endosperm type Normal starch Weight per 100 kernels  28.1 gm #9Cob Diameter at mid-point  28.2 mm Cob color 13 cherry red (Munsell Code10R 5/8) *scale from 1 = none to 9 = like peach fuzz **scale from 1-noneto 9-many ***scale from 0 = male sterile to 9 = heavy shed ****scalefrom 1 to 9; 1 = loose 9 = tightHeat Units per day were calculated: HU=[MaxTemp (86)=Min Temp(50)]/2-50. Large standard deviations are probable due to environmentalfactors at the locations of observation. Inbred's response toenvironment is often more pronounced than a hybrid.

The comparable inbred to G06-NP2546 is MO17. The present invention hasdiffering ear leaf length and a significantly smaller ear size than doesA619. Other traits that differ are the cob diameter and the ear diameterand the ear length which is shorter for the present invention.Additionally, the present invention is also an earlier flowering inbredthan is A619.

The data provided above is often a color. The Munsell code is areference book of color, which is known and used in the industry and bypersons with ordinary skill in the art of plant breeding.

Inbred Performance of G06-NP2546

The traits and characteristics of inbred corn line G06-NP2546 are listedto compare with other inbreds. The G06-NP2546 data shows thecharacteristics and traits of importance, giving a snapshot ofG06-NP2546 in these specific environments.

TABLE 2 PAIRED INBRED COMPARISON DATA Heat Heat Units Cob Ear Ear LeafEar Heat Units Units to 90% Leaf Tassel Diameter Diameter Length Lengthto 10% to 50% Pollen Sheath Length (mm) (mm) (cm) (cm) Pollen Shed SilkShed Pubscence (cm) G06- 24 39 81 12 986 1009 1073 7 33 NP2536 A619 2644 70 15 1032 1095 1156 2 45 Mean 25 42 76 13 1009 1052 1110 5 39General Trials 5 5 5 5 5 5 5 5 5 w/data Entries 2 2 2 2 2 2 2 2 2 w/dataLSD 1 2 6 2 40 73 67 2 5 General (95% confidence level) CV 2 2 4 8 2 4 326 8 (Effective) % Probability 1 0 1 2 3 3 3 0 0 %The cob and ear of the present invention and that of Mo17 are different.The cob girth of the present invention is larger than Mo17. Likewise theear girth of the present invention is larger than Mo17. The Ear leafLength is also different as the present inventions is shorter.The present invention is earlier to silk and is faster to 90% pollenshed than is Mo17. The anther colors show a strong difference betweenthe two lines also.The present invention is shown in Table 2 is the mean from individualdata gathered for two years at two separate locations. This data wasgathered to produce a snapshot of the inbred across a number of physicaland environmental conditions. This data shows an inbred that has asmaller cob size than does the comparison inbred.

This invention also is directed to methods for producing a corn plant bycrossing a first parent corn plant with a second parent corn plantwherein the first or second parent corn plant is an inbred corn plantfrom the line G06-NP2546. Further, both first and second parent cornplants can come from the inbred corn line G06-NP2546 which produces aself of the inbred invention. The present invention can be employed in avariety of breeding methods which can be selected depending on the modeof reproduction, the trait, and the condition of the germplasm. Thus,any breeding methods using the inbred corn line G06-NP2546 are part ofthis invention: selfing, backcrosses, hybrid production, crosses topopulations, haploid by such old and known methods of using stock sixmaterial that induces haploids and anther culturing and the like.

All plants and plant cells produced using inbred corn line G06-NP2546are within the scope of this invention. The invention encompasses theinbred corn line used in crosses with other, different, corn inbreds toproduce (F1) corn hybrid seeds and hybrid plants and the grain producedon the hybrid plant. This invention includes plant and plant cells,which upon growth and differentiation produce corn plants having thephysiological and morphological characteristics of the inbred lineG06-NP2546.

Additionally, this maize can, within the scope of the invention,contain: a mutant gene such as, but not limited to, the amylose,amylase, sugary 1, shrunken 1, waxy, AE or imazethapyr tolerant (IT orIR™) mutant gene; or transgenic genes such as but not limited to insectresistant genes such as Corn Rootworm genes, Bacillus thuringiensis (Crygenes), or herbicide resistant genes such as Pat gene or Bar gene, EPSP,GOX, Glyphosate resistant genes, or disease resistant genes such as theMosaic virus resistant gene, etc., or trait altering genes such asflowering genes, oil modifying genes, senescence genes and the like. Themethods and techniques for inserting, or producing and/or identifying amutation or a transgene into the present invention through breeding,transformation, or mutating are well known and understood by those ofordinary skill in the art.

Various techniques for breeding and moving or altering genetic materialwithin or into the present invention (whether it is an inbred or inhybrid combination) are also known to those skilled in the art. Thesetechniques to list only a few are anther culturing, haploid production,(stock six is a method that has been in use for thirty years and is wellknown to those with skill in the art), transformation, irradiation toproduce mutations, chemical or biological mutation agents and a host ofother methods are within the scope of the invention. All parts of theG06-NP2546 plant including its plant cells produced using the inbredcorn line is within the scope of this invention. The term transgenicplant refers to plants having genetic sequences, which are introducedinto the genome of a plant by a transformation method and the progenythereof. Transformation methods are means for integrating new geneticcoding sequences into the plant's genome by the incorporation of thesesequences into a plant through man's assistance, but not by breedingpractices. The transgene once introduced into plant material andintegrated stably can be moved into other germplasm by standard breedingpractices.

Though there are a large number of known methods to transform plants,certain types of plants are more amenable to transformation than areothers. Transformation of dicots is usually achievable for example,tobacco is a readily transformable plant. Monocots can present sometransformation challenges, however, the basic steps of transformingplants monocots have been known in the art for about 15 years. The mostcommon method of maize transformation is referred to as gunning ormicroprojectile bombardment though other methods can be used. Theprocess employs small gold-coated particles coated with DNA which areshot into the transformable material. Detailed techniques for gunningDNA into cells, tissue, callus, embryos, and the like are well known inthe prior art. One example of steps that can be involved in monocottransformation are concisely outlined in U.S. Pat. No. 5,484,956“Fertile Transgenic Zea mays Plants Comprising Heterologous DNA EncodingBacillus Thuringiensis Endotoxin” issued Jan. 16, 1996 and also in U.S.Pat. No. 5,489,520 “Process of Producing Fertile Zea mays Plants andProgeny Comprising a Gene Encoding Phosphinothricin Acetyl Transferase”issued Feb. 6, 1996.

Plant cells such as maize can be transformed not only by the use of agunning device but also by a number of different techniques. Some ofthese techniques include maize pollen transformation (See University ofToledo 1993 U.S. Pat. No. 5,177,010); Whiskers technology (See U.S. Pat.Nos. 5,464,765 and 5,302,523); electroporation; PEG on Maize;Agrobacterium (See 1996 article on transformation of maize cells inNature Biotechnology, Volume 14, June 1996) along with numerous othermethods which may have slightly lower efficiency rates. Some of thesemethods require specific types of cells and other methods can bepracticed on any number of cell types.

The use of pollen, cotyledons, zygotic embryos, meristems and ovum asthe target issue can eliminate the need for extensive tissue culturework. Generally, cells derived from meristematic tissue are useful. Themethod of transformation of meristematic cells of cereal is taught inthe PCT application WO96/04392. Any number of various cell lines,tissues, calli and plant parts can and have been transformed by thosehaving knowledge in the art. Methods of preparing callus or protoplastsfrom various plants are well known in the art and specific methods aredetailed in patents and references used by those skilled in the art.Cultures can be initiated from most of the above-identified tissue. Theonly true requirement of the transforming plant material is that it canform a transformed plant.

The DNA used for transformation of these plants clearly may be circular,linear, and double or single stranded. Usually, the DNA is in the formof a plasmid. The plasmid usually contains regulatory and/or targetingsequences which assists the expression of the gene in the plant. Themethods of forming plasmids for transformation are known in the art.Plasmid components can include such items as: leader sequences, transitpolypeptides, promoters, terminators, genes, introns, marker genes, etc.The structures of the gene orientations can be sense, antisense, partialantisense, or partial sense: multiple gene copies can be used. Thetransgenic gene can come from various non-plant genes (such as;bacteria, yeast, animals, and viruses) along with being from plants.

The regulatory promoters employed can be constitutive such as CaMv35S(usually for dicots) and polyubiquitin for monocots or tissue specificpromoters such as CAB promoters, MR7 described in U.S. Pat. No.5,837,848, etc. The prior art promoters, includes but is not limited to,octopine synthase, nopaline synthase, CaMv19S, mannopine synthase. Theseregulatory sequences can be combined with introns, terminators,enhancers, leader sequences and the like in the material used fortransformation.

The isolated DNA is then transformed into the plant. After thetransformation of the plant material is complete, the next step isidentifying the cells or material, which has been transformed. In somecases, a screenable marker is employed such as the beta-glucuronidasegene of the uidA locus of E. coli. Then, the transformed cellsexpressing the colored protein are selected. In many cases, a selectablemarker identifies the transformed material. The putatively transformedmaterial is exposed to a toxic agent at varying concentrations. Thecells not transformed with the selectable marker, which providesresistance to this toxic agent, die. Cells or tissues containing theresistant selectable marker generally proliferate. It has been notedthat although selectable markers protect the cells from some of thetoxic affects of the herbicide or antibiotic, the cells may still beslightly affected by the toxic agent by having slower growth rates. Ifthe transformed material was cell lines then these lines are regeneratedinto plants. The cells' lines are treated to induce tissuedifferentiation. Methods of regeneration of cellular maize material arewell known in the art.

A deposit of at least 2500 seeds of this invention will be maintained bySyngenta Seed Inc. Access to this deposit will be available during thependency of this application to the Commissioner of Patents andTrademarks and persons determined by the Commissioner to be entitledthereto upon request. All restrictions on availability to the public ofsuch material will be removed upon issuance of a granted patent of thisapplication by depositing at least 2500 seeds of this invention at theAmerican Type Culture Collection (ATCC), at 10801 University Boulevard,Manassas, Va. 20110. The date of deposit was Feb. 8, 2008. The ATCCnumber of the deposit is PTA-8934 and on Feb. 29, 2008 the deposit wasfound viable when tested. The deposit of at least 2500 seeds will befrom the same inbred seed taken from the deposit maintained by SyngentaSeed Inc. The ATCC deposit will be maintained in that depository, whichis a public depository, for a period of 30 years, or 5 years after thelast request, or for the enforceable life of the patent, whichever islonger, and will be replaced if it becomes nonviable during that period.

Additional public information on the present invention may be availablefrom the PVP Office, a division of the US Government.

Accordingly, the present invention has been described with some degreeof particularity directed to the preferred embodiment of the presentinvention. It should be appreciated, though, that the present inventionis defined by the following claims construed in light of the prior artso that modifications or changes may be made to the preferred embodimentof the present invention without departing from the inventive conceptscontained herein.

1. Seed of maize inbred line designated G06-NP2546, representative seedof said line having been deposited under ATCC Accession No. PTA-8934. 2.A maize plant, or a part thereof, produced by growing the seed ofclaim
 1. 3. The maize plant of claim 2, wherein said plant has beendetasseled.
 4. A tissue culture of regenerable cells produced from theplant of claim
 2. 5. Protoplasts produced from the tissue culture ofclaim
 4. 6. The tissue culture of claim 4, wherein cells of the tissueculture are from a plant part selected from the group consisting ofleaf, pollen, embryo, root, root tip, anther, silk, flower, kernel, ear,cob, husk and stalk.
 7. A maize plant regenerated from the tissueculture of claim 4, said plant having all the morphological andphysiological characteristics of inbred line G06-NP2546, representativeseed of said line having been deposited under ATCC Accession No.PTA-8934.
 8. A method for producing an F1 hybrid maize seed, said methodcomprising crossing the plant of claim 2 with a different maize plantand harvesting the resultant F1 hybrid maize seed.
 9. A method ofproducing a male sterile maize plant, said method comprisingtransforming the maize plant of claim 2 with a nucleic acid moleculethat confers male sterility.
 10. A male sterile maize plant produced bythe method of claim
 9. 11. A method of producing an herbicide resistantmaize plant, said method comprising transforming the maize plant ofclaim 2 with a transgene that confers herbicide resistance.
 12. Anherbicide resistant maize plant produced by the method of claim
 11. 13.The maize plant of claim 12, wherein the transgene confers resistance toan herbicide selected from the group consisting of: imidazolinone,sulfonylurea, glyphosate, glufosinate, L-phosphinothricin, triazine andbenzonitrile.
 14. A method of producing an insect resistant maize plant,said method comprising transforming the maize plant of claim 2 with atransgene that confers insect resistance.
 15. An insect resistant maizeplant produced by the method of claim
 14. 16. The maize plant of claim15, wherein the transgene encodes a Bacillus thuringiensis endotoxin.17. A method of producing a disease resistant maize plant, said methodcomprising transforming the maize plant of claim 2 with a transgene thatconfers disease resistance.
 18. A disease resistant maize plant producedby the method of claim
 17. 19. A method of producing a maize plant withdecreased phytate content, said method comprising transforming the maizeplant of claim 2 with a transgene encoding phytase.
 20. A maize plantwith decreased phytate content produced by the method of claim
 19. 21. Amethod of producing a maize plant with modified fatty acid metabolism ormodified carbohydrate metabolism, said method comprising transformingthe maize plant of claim 2 with a transgene encoding a protein selectedfrom the group consisting of stearyl-ACP desaturase,fructosyltransferase, levansucrase, alpha-amylase, invertase and starchbranching enzyme.
 22. A maize plant produced by the method of claim 21.23. The maize plant of claim 22, wherein the transgene confers a traitselected from the group consisting of waxy starch and increased amylosestarch.
 24. A maize plant, or part thereof, having all the physiologicaland morphological characteristics of the inbred line G06-NP2546,representative seed of said line having been deposited under ATCCAccession No. PTA-8934.
 25. A method of introducing a desired trait intomaize inbred line G06-NP2546, said method comprising: (a) crossingG06-NP2546 plants grown from G06-NP2546 seed, representative seed ofwhich has been deposited under ATCC Accession No. PTA-8934, with plantsof another maize line that comprise a desired trait to produce F1progeny plants, wherein the desired trait is selected from the groupconsisting of male sterility, herbicide resistance, insect resistance,disease resistance and waxy starch; (b) selecting F1 progeny plants thathave the desired trait to produce selected F1 progeny plants; (c)crossing the selected progeny plants with the G06-NP2546 plants toproduce backcross progeny plants; (d) selecting for backcross progenyplants that have the desired trait and physiological and morphologicalcharacteristics of maize inbred line G06-NP2546 listed in Table 1 toproduce selected backcross progeny plants; and (e) repeating steps (c)and (d) three or more times in succession to produce selected fourth orhigher backcross progeny plants that comprise the desired trait and allof the physiological and morphological characteristics of maize inbredline G06-NP2546 listed in Table 1 as determined at the 5% significancelevel when grown in the same environmental conditions.
 26. A plantproduced by the method of claim 25, wherein the plant has the desiredtrait and all of the physiological and morphological characteristics ofmaize inbred line G06-NP2546 listed in Table 1 as determined at the 5%significance level when grown in the same environmental conditions. 27.The plant of claim 26, wherein the desired trait is herbicide resistanceand the resistance is conferred to an herbicide selected from the groupconsisting of: imidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine and benzonitrile.
 28. The plant of claim26, wherein the desired trait is insect resistance and the insectresistance is conferred by a transgene encoding a Bacillus thuringiensisendotoxin.
 29. The plant of claim 26, wherein the desired trait is malesterility and the trait is conferred by a cytoplasmic nucleic acidmolecule that confers male sterility.
 30. A method of modifying fattyacid metabolism, modified phytic acid metabolism or modifiedcarbohydrate metabolism into maize inbred line G06-NP2546, said methodcomprising: (a) crossing G06-NP2546 plants grown from G06-NP2546 seed,representative seed of which has been deposited under ATCC Accession No.PTA-8934, with plants of another maize line that comprise a nucleic acidmolecule encoding an enzyme selected from the group consisting ofphytase, stearyl-ACP desaturase, fructosyltransferase, levansucrase,alphaamylase, invertase and starch branching enzyme; (b) selecting F1progeny plants that have said nucleic acid molecule to produce selectedF1 progeny plants; (c) crossing the selected progeny plants with theG06-NP2546 plants to produce backcross progeny plants; (d) selecting forbackcross progeny plants that have said nucleic acid molecule andphysiological and morphological characteristics of maize inbred lineG06-NP2546 listed in Table 1 to produce selected backcross progenyplants; and (e) repeating steps (c) and (d) three or more times insuccession to produce selected fourth or higher backcross progeny plantsthat comprise said nucleic acid molecule and have all of thephysiological and morphological characteristics of maize inbred lineG06-NP2546 listed in Table 1 as determined at the 5% significance levelwhen grown in the same environmental conditions.
 31. A plant produced bythe method of claim 30, wherein the plant comprises the nucleic acidmolecule and has all of the physiological and morphologicalcharacteristics of maize inbred line G06-NP2546 listed in Table 1 asdetermined at the 5% significance level when grown in the sameenvironmental conditions.